The present invention relates to novel classes of compounds which are caspase inhibitors, in particular interleukin-1xcex2 converting enzyme (xe2x80x9cICExe2x80x9d) inhibitors. This invention also relates to pharmaceutical compositions comprising these compounds. The compounds and pharmaceutical compositions of this invention are particularly well suited for inhibiting caspase activity and consequently, may be advantageously used as agents against interleukin-1-(xe2x80x9cIL-1xe2x80x9d), apoptosis-, interferon-xcex3 inducing factor-(IGIF), or interferon-(xe2x80x9cIFN-xcex3xe2x80x9d) mediated diseases, including inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, and degenerative diseases. This invention also relates to methods for inhibiting caspase activity and decreasing IGIF production and IFN-xcex3 production and methods for treating interleukin-1, apoptosis-, and interferon-xcex3-mediated diseases using the compounds and compositions of this invention. This invention also relates to methods of preparing the compounds of this invention.
Interleukin 1 (xe2x80x9cIL-1xe2x80x9d) is a major pro-inflammatory and immunoregulatory protein that stimulates fibroblast differentiation and proliferation, the production of prostaglandins, collagenase and phospholipase by synovial cells and chondrocytes, basophil and eosinophil degranulation and neutrophil activation. Oppenheim, J. H. et al, Immunology Today, 7, pp. 45-56 (1986). As such, it is involved in the pathogenesis of chronic and acute inflammatory and autoimmune diseases. For example, in rheumatoid arthritis, IL-1 is both a mediator of inflammatory symptoms and of the destruction of the cartilage proteoglycan in afflicted joints. Wood, D. D. et al., Arthritis Rheum. 26, 975, (1983); Pettipher, E. J. et al., Proc. Natl. Acad. Sci. USA 71, 295 (1986); Arend, W. P. and Dayer, J. M., Arthritis Rheum. 38, 151 (1995). IL-1 is also a highly potent bone resorption agent. Jandiski, J. J., J. Oral Path 17, 145 (1988); Dewhirst, F. E. et al., J. Immunol. 8, 2562 1985). It is alternately referred to as xe2x80x9costeoclast activating factorxe2x80x9d in destructive bone diseases such as osteoarthritis and multiple myeloma. Bataille, R. et al., Int. J. Clin. Lab. Res. 21(4), 283 (1992). In certain proliferative disorders, such as acute myelogenous leukemia and multiple myeloma, IL-1 can promote tumor cell growth and adhesion. Bani, M. R., J. Natl. Cancer Inst. 83, 123 (1991); Vidal-Vanaclocha, F., Cancer Res. 54, 2667 (1994). In these disorders, IL-1 also stimulates production of other cytokines such as IL-6, which can modulate tumor development (Tartour et al., Cancer Res. 54, p. 6243 (1994). IL-1 is predominantly produced by peripheral blood monocytes as part of the inflammatory response and exists in two distinct agonist forms, IL-1xcex1 and IL-1xcex2. Mosely, B. S. et al., Proc. Nat. Acad. Sci., 84, pp. 4572-4576 (1987); Lonnemann, G. et al., Eur. J. Immunol., 19, pp. 1531-1536 (1989).
IL-1xcex2 is synthesized as a biologically inactive precursor, pIL-1xcex2. pIL-1xcex2 lacks a conventional leader sequence and is not processed by a signal peptidase. March, C. J., Nature, 315, pp. 641-647 (1985). Instead, pIL-1xcex2 is cleaved by interleukin-1xcex2 converting enzyme (xe2x80x9cICExe2x80x9d) between Asp-116 and Ala-117 to produce the biologically active C-terminal fragment found in human serum and synovial fluid. Sleath, P. R., et al., J. Biol. Chem., 265, pp. 14526-14528 (1992); A. D. Howard et al., J. Immunol., 147, pp. 2964-2969 (1991). ICE is a cysteine protease localized primarily in monocytes. It converts precursor IL-1xcex2 to the mature form. Black, R. A. et al., FEBS Lett., 247, pp. 386-390 (1989); Kostura, M. J. et al., Proc. Natl. Acad. Sci. U.S.A., 86, pp. 5227-5231 (1989). Processing by ICE is also necessary for the transport of mature IL-1xcex2 through the cell membrane.
ICE (or caspase-1) is a member of a family of homologous enzymes called caspases. These homologs have sequence similarities in the active site regions of the enzymes. Such homologs (caspases) include TX (or ICErel-II or ICH-2) (caspase-4) (Faucheu, et al., EMBO J., 14, p. 1914 (1995); Kamens J., et al., J. Biol. Chem., 270, p. 15250 (1995); Nicholson et al., J. Biol. Chem., 270 15870 (1995)), TY (or ICErel-III) (caspase-5) (Nicholson et al., J. Biol. Chem., 270, p. 15870 (1995); ICH-1 (or Nedd-2) (caspase-2) (Wang, L. et al., Cell, 78, p. 739 (1994)), MCH-2 (caspase-6), (Fernandes-Alnemri, T. et al., Cancer Res., 55, p. 2737 (1995), CPP32 (or YAMA or apopain) (caspase-3) (Fernandes-Alnemri, T. et al., J. Biol. Chem., 269, p. 30761 (1994); Nicholson, D. W. et al., Nature, 376, p. 37 (1995)), CMH-1 (or MCH-3) (caspase-7) (Lippke, et al., J. Biol. Chem., 271(4), p 1825-1828 (1996)); Fernandes-Alnemri, T. et al., Cancer Res., (1995)), Mch5 (caspase-8) (Muzio, M. et. al., Cell 85(6), 817-827, (1996)), MCH-6 (caspase-9) (Duan, H. et. al., J. Biol. Chem., 271(34), p. 16720-16724 (1996)), Mch4 (caspase-10) (Vincenz, C. et. al., J. Biol. Chem., 272, p. 6578-6583 (1997); Fernandes-Alnemri, T. et. al., Proc. Natl. Acad. Sci. 93, p. 7464-7469 (1996)), Ich-3 (caspase-11) (Wang, S. et. al., J. Biol. Chem., 271, p. 20580-20587 (1996)), mCASP-12 (caspase-12), (Van de Craen, M. et. al., FEBS Lett. 403, p. 61-69 (1997); Yuan, Y. and Miura, M. PCT Publication WO95/00160 (1995)), ERICE (caspase-13), (Humke, E. W., et. al., J. Biol. Chem., 273(25) p. 15702-15707 (1998)), and MICE (caspase-14) (Hu, S. et. al., J. Biol. Chem., 273(45) p. 29648-29653 (1998)).
Each of these ICE homologs, as well as ICE itself, is capable of inducing apoptosis when overexpressed in transfected cell lines. Inhibition of one or more of these homologs with the peptidyl ICE inhibitor Tyr-Val-Ala-Asp-chloromethylketone results in inhibition of apoptosis in primary cells or cell lines. Lazebnik et al., Nature, 371, p. 346 (1994).
Caspases also appear to be involved in the regulation of programmed cell death or apoptosis. Yuan, J. et al., Cell, 75, pp. 641-652 (1993); Miura, M. et al., Cell, 75, pp. 653-660 (1993); Nett-Fiordalisi, M. A. et al., J. Cell Biochem., 17B, p. 117 (1993). In particular, ICE or ICE homologs are thought to be associated with the regulation of apoptosis in neurodegenerative diseases, such as Alzheimer""s and Parkinson""s disease. Marx, J. and M. Baringa, Science, 259, pp. 760-762 (1993); Gagliardini, V. et al., Science, 263, pp. 826-828 (1994). Therapeutic applications for inhibition of apoptosis may include treatment of Alzheimer""s disease, Parkinson""s disease, stroke, myocardial infarction, spinal atrophy, and aging.
ICE has been demonstrated to mediate apoptosis (programmed cell death) in certain tissue types. Steller, H., Science, 267, p. 1445 (1995); Whyte, M. and Evan, G., Nature, 376, p. 17 (1995); Martin, S. J. and Green, D. R., Cell, 82, p. 349 (1995); Alnemri, E. S., et al., J. Biol. Chem., 270, p. 4312 (1995); Yuan, J. Curr. Opin. Cell Biol., 7, p. 211 (1995). A transgenic mouse with a disruption of the ICE gene is deficient in Fas-mediated apoptosis (Kuida, K. et al., Science 267, 2000 (1995)). This activity of ICE is distinct from its role as the processing enzyme for pro-IL-1xcex2. It is conceivable that in certain tissue types, inhibition of ICE may not affect secretion of mature IL-1xcex2, but may inhibit apoptosis.
Enzymatically active ICE has been previously described as a heterodimer composed of two subunits, p20 and p10 (20 kDa and 10 kDa molecular weight, respectively). These subunits are derived from a 45 kDa proenzyme (p45) by way of a p30 form, through an activation mechanism that is autocatalytic. Thornberry, N. A. et al., Nature, 356, pp. 768-774 (1992). The ICE proenzyme has been divided into several functional domains: a prodomain (p14), a p22/20 subunit, a polypeptide linker and a p10 subunit. Thornberry et al., supra; Casano et al., Genomics, 20, pp. 474-481 (1994).
Full length p45 has been characterized by its cDNA and amino acid sequences. PCT patent applications WO 91/15577 and WO 94/00154. The p20 and p10 cDNA and amino acid sequences are also known. Thornberry et al., supra. Murine and rat ICE have also been sequenced and cloned. They have high amino acid and nucleic acid sequence homology to human ICE. Miller, D. K. et al., Ann. N.Y. Acad. Sci., 696, pp. 133-148 (1993); Molineaux, S. M. et al., Proc. Nat. Acad. Sci., 90, pp. 1809-1813 (1993). The three-dimensional structure of ICE has been determined at atomic resolution by X-ray crystallography. Wilson, K. P., et al., Nature, 370, pp. 270-275 (1994). The active enzyme exists as a tetramer of two p20 and two p10 subunits.
Recently, ICE and other members of the ICE/CED-3 family have been linked to the conversion of pro-IGIF to IGIF or to the production of IFN-xcex3 in vivo (PCT application PCT/US96/20843, publication no. WO 97/22619, which is incorporated herein by reference). IGIF is synthesized in vivo as the precursor protein xe2x80x9cpro-IGIFxe2x80x9d.
Interferon-gamma inducing factor (IGIF) is an approximately 18-kDa polypeptide that stimulates T-cell production of interferon-gamma (IFN-xcex3). IGIF is produced by activated Kupffer cells and macrophages in vivo and is exported out of such cells upon endotoxin stimulation. Thus, a compound that decreases IGIF production would be useful as an inhibitor of such T-cell stimulation which in turn would reduce the levels of-IFN-xcex3 production by those cells.
IFN-xcex3 is a cytokine with immunomodulatory effects on a variety of immune cells. In particular, IFN-xcex3 is involved in macrophage activation and Th1 cell selection (F. Belardelli, APMIS, 103, p. 161 (1995)). IFN-xcex3 exerts its effects in part by modulating the expression of genes through the STAT and IRF pathways (C. Schindler and J. E. Darnell, Ann. Rev. Biochem., 64, p. 621 (1995); T. Taniguchi, J. Cancer Res. Clin. Oncol., 121, p. 516 (1995)).
Mice lacking IFN-xcex3 or its receptor have multiple defects in immune cell function and are resistant to endotoxic shock (S. Huang et al., Science, 259, p. 1742 (1993); D. Dalton et al., Science, 259, p. 1739 (1993); B. D. Car et al., J. Exp. Med., 179, p. 1437 (1994)). Along with IL-12, IGIF appears to be a potent inducer of IFN-xcex3 production by T cells (H. Okamura et al., Infection and Immunity, 63, p. 3966 (1995); H. Okamura et al., Nature, 378, p. 88 (1995); S. Ushio et al., J. Immunol., 156, p. 4274 (1996)).
IFN-xcex3 has been shown to contribute to the pathology associated with a variety of inflammatory, infectious and autoimmune disorders and diseases. Thus, compounds capable of decreasing IFN-xcex3 production would be useful to ameliorate the effects of IFN-xcex3 related pathologies.
Accordingly, compositions and methods capable of regulating the conversion of pro-IGIF to IGIF would be useful for decreasing IGIF and IFN-xcex3 production in vivo, and thus for ameliorating the detrimental effects of these proteins which contribute to human disorders and diseases.
Caspase inhibitors represent a class of compounds useful for the control of inflammation or apoptosis or both. Peptide and peptidyl inhibitors of ICE have been described (PCT patent applications WO 91/15577, WO 93/05071, WO 93/09135, WO 93/12076, WO 93/14777, WO 93/16710, WO 95/35308, WO 96/30395, WO 96/33209 and WO 98/01133; European patent applications 503 561, 547 699, 618 223, 623 592, and 623 606; and U.S. Pat. Nos. 5,434,248, 5,710,153, 5,716,929, and 5,744,451). Such peptidyl inhibitors of ICE have been observed to block the production of mature IL-1xcex2 in a mouse model of inflammation (vide infra) and to suppress growth of leukemia cells in vitro (Estrov et al., Blood, 84, 380a (1994)). However, due to their peptidic nature, such inhibitors are typically characterized by undesirable pharmacologic properties, such as poor cellular penetration and cellular activity, poor oral absorption, instability and rapid metabolism. Plattner, J. J. and D. W. Norbeck, in Drug Discovery Technologies, C. R. Clark and W. H. Moos, Eds. (Ellis Horwood, Chichester, England, 1990), pp. 92-126. These properties has hampered their development into effective drugs.
Non-peptidyl compounds have also been reported to inhibit ICE in vitro. PCT patent application WO 95/26958; U.S. Pat. No. 5,552,400; Dolle et al., J. Med. Chem., 39, pp. 2438-2440 (1996).
It is not clear however whether these compounds have the appropriate pharmacological profiles to be therapeutically useful.
Accordingly, the need exists for compounds that can effectively inhibit caspases, and that have favorable in vivo activity, for use as agents for preventing and treating chronic and acute forms of IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated diseases, as well as inflammatory, autoimmune, destructive bone, proliferative, infectious, or degenerative diseases.
The present invention provides novel classes of compounds, and pharmaceutically acceptable derivatives thereof, that are useful as caspase inhibitors, in particular, as ICE inhibitors. These compounds can be used alone or in combination with other therapeutic or prophylactic agents, such as antibiotics, immunomodulators or other anti-inflammatory agents, for the treatment or prophylaxis of diseases mediated by IL-1, apoptosis, IGIF, or IFN-xcex3. According to a preferred embodiment, the compounds of this invention are capable of binding to the active site of a caspase and inhibiting the activity of that enzyme.
It is a principal object of this invention to provide novel classes of compounds represented by formula I, which have favorable in vivo profiles: 
wherein the various substituents are described herein.
It is a further objective of this invention to provide pharmaceutical compositions, including multi-component compositions. This invention also provides methods for using and preparing the compounds of this invention and related compounds.
In order that the invention described herein may be more fully understood, the following detailed description is set forth.
The following abbreviations and definitions are used throughout the application.
The term xe2x80x9ccaspasexe2x80x9d refers to an enzyme that is a member of the family of enzymes that includes ICE (see H. Hara, Natl. Acad. Sci., 94, pp. 2007-2012 (1997)).
The terms xe2x80x9cHBVxe2x80x9d, xe2x80x9cHCVxe2x80x9d and xe2x80x9cHGVxe2x80x9d refer to hepatitis-B virus, hepatitis-C virus and hepatitis-G virus, respectively.
The term xe2x80x9cKixe2x80x9d refers to a numerical measure of the effectiveness of a compound in inhibiting the activity of a target enzyme such as ICE. Lower values of Ki reflect higher effectiveness. The Ki value is a derived by fitting experimentally determined rate data to standard enzyme kinetic equations (see I. H. Segel, Enzyme Kinetics, Wiley-Interscience, 1975).
The term xe2x80x9cinterferon gamma inducing factorxe2x80x9d or xe2x80x9cIGIFxe2x80x9d refers to a factor which is capable of stimulating the endogenous production of IFN-xcex3.
The term xe2x80x9ccaspase inhibitorxe2x80x9d refer to a compound which is capable of demonstrating detectable inhibition of one or more caspases. The term xe2x80x9cICE inhibitorxe2x80x9d refers to a compound which is capable of demonstrating detectable inhibition of ICE and optionally one or more additional caspases. Inhibition of these enzymes may be determined using the methods described and incorporated by reference herein.
The skilled practitioner realizes that an in vivo enzyme inhibitor is not necessarily an in vitro enzyme inhibitor. For example, a prodrug form of a compound typically demonstrates little or no activity in in vitro assays. Such prodrug forms may be altered by metabolic or other biochemical processes in the patient to provide an in vivo ICE inhibitor.
The term xe2x80x9ccytokinexe2x80x9d refers to a molecule which mediates interactions between cells.
The term xe2x80x9cconditionxe2x80x9d refers to any disease, disorder or effect that produces deleterious biological consequences in a subject.
The term xe2x80x9csubjectxe2x80x9d refers to an animal, or to one or more cells derived from an animal. Preferably, the animal is a mammal, most preferably a human. Cells may be in any form, including but not limited to cells retained in tissue, cell clusters, immortalized cells, transfected or transformed cells, and cells derived from an animal that have been physically or phenotypically altered.
The term xe2x80x9cpatientxe2x80x9d as used in this application refers to any mammal, preferably humans.
The term xe2x80x9calkylxe2x80x9d refers to a straight-chained or branched, saturated aliphatic hydrocarbon containing 1 to 6 carbon atoms.
The term xe2x80x9calkenylxe2x80x9d refers to a straight-chained or branched unsaturated hydrocarbon containing 2 to 6 carbon atoms and at least one double bond.
The term xe2x80x9calkynylxe2x80x9d refers to a straight-chained or branched unsaturated hydrocarbon containing 2 to 6 carbon atoms and at least one triple bond.
The term xe2x80x9ccycloalkylxe2x80x9d refers to a mono- or polycyclic, non-aromatic, hydrocarbon ring system which may optionally contain unsaturated bonds in the ring system. Examples include cyclohexyl, adamantyl, norbornyl, and spirocyclopentyl.
The term xe2x80x9carylxe2x80x9d refers to a mono- or polycyclic ring system which contains 6, 10, 12 or 14 carbons in which at least one ring of the ring system is aromatic. The aryl groups of this invention are optionally singly or multiply substituted with R11. Examples of aryl ring systems include, phenyl, naphthyl, and tetrahydronaphthyl.
The term xe2x80x9cheteroarylxe2x80x9d refers to a mono- or polycyclic ring system which contains 1 to 15 carbon atoms and 1 to 4 heteroatoms, and in which at least one ring of the ring system is aromatic. Heteroatoms are sulfur, nitrogen or oxygen. The heteroaryl groups of this invention are optionally singly or multiply substituted with R11.
The term xe2x80x9cheterocyclicxe2x80x9d refers to a mono- or polycyclic ring system which contains 1 to 15 carbon atoms and 1 to 4 heteroatoms, in which the mono- or polycyclic ring system may optionally contain unsaturated bonds but is not aromatic. Heteroatoms are independently sulfur, nitrogen, or oxygen.
The term xe2x80x9calkylarylxe2x80x9d refers to an alkyl group, wherein a hydrogen atom of the alkyl group is replaced by an aryl radical.
The term xe2x80x9calkylheteroarylxe2x80x9d refers to an alkyl group, wherein a hydrogen atom of the alkyl group is replaced by a heteroaryl radical.
The term xe2x80x9camino acid side chainxe2x80x9d refers to any group attached to the a carbon of a naturally or non-naturally occuring amino acid.
The term xe2x80x9csubstitutexe2x80x9d refers to the replacement of a hydrogen atom in a compound with a substituent group.
The term xe2x80x9cstraight chainxe2x80x9d refers to a contiguous unbranching string of covalently bound atoms. The straight chain may be substituted, but these substituents are not a part of the straight chain.
In chemical formulas, parenthesis are used herein to denote connectivity in molecules or groups. In particular, parentheses are used to indicate: 1) that more than one atom or group is bonded to a particular atom; or 2) a branching point (i.e., the atom immediately before the open parenthesis is bonded both to the atom or group in the parentheses and the atom or group immediately after the closed parenthesis). An example of the first use is xe2x80x9cxe2x80x94N(alkyl)2xe2x80x9d, indicating two alkyl groups bond to an N atom. An example of the second use is xe2x80x9cxe2x80x94C(O)NH2xe2x80x9d, indicating a carbonyl group and an amino (xe2x80x9cNH2xe2x80x9d) group, both bonded to the indicated carbon atom. A xe2x80x9cxe2x80x94C(O)NH2xe2x80x9d group may be represented in other ways, including the following structure: 
Substituents may be represented in various forms. These various forms are known to the skilled practitioner and may be used interchangeably. For example, a methyl substituent on a phenyl ring may be represented in any of the following forms: 
Various forms of substituents such as methyl are used herein interchangeably.
Other definitions are set forth in the specification where necessary.
The compounds of one embodiment A of this invention are those of formula I: 
wherein:
Y is: 
xe2x80x83provided that when R7 is xe2x80x94OH then Y can also be: 
X is xe2x80x94C(R3)2xe2x80x94 or xe2x80x94N(R3)xe2x80x94;
m is 0 or 1;
R1 is H, xe2x80x94C(O)R8, xe2x80x94C(O)C(O)R8, xe2x80x94S(O)2R8, xe2x80x94S(O)R8, xe2x80x94C(O)OR8, xe2x80x94C(O)N(H)R8, xe2x80x94S(O)2N(H)xe2x80x94R8, xe2x80x94S(O)N(H)xe2x80x94R8, xe2x80x94C(O)C(O)N(H)R8, xe2x80x94C(O)CHxe2x95x90CHR8, xe2x80x94C(O)CH2OR8, xe2x80x94C(O)CH2N(H)R8, xe2x80x94C(O)N(R8)2, xe2x80x94S(O)2N(R8)2, xe2x80x94S(O)N(R8)2, xe2x80x94C(O)C(O)N(R8)2, xe2x80x94C(O)CH2N(R8)2, xe2x80x94CH2R8, xe2x80x94CH2-alkenyl-R8, or xe2x80x94CH2-alkynyl-R8;
R2 is xe2x80x94H and each R3 is independently xe2x80x94H, an amino acid side chain, xe2x80x94R8, alkenyl-R9, or alkynyl-R9, or R2 and one R3 together with the atoms to which they are bound, form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by xe2x80x94R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by xe2x80x94R11, a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced by xe2x80x94R1;
R4 is xe2x80x94H and each R5 is independently xe2x80x94H, an amino acid side chain, xe2x80x94R8, -alkenyl-R9, or -alkynyl-R9, or R4 and one R5 together with the atoms to which they are bound form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R1, and a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced with R1;
R6 is xe2x80x94H;
R7 is xe2x80x94OH, xe2x80x94OR8, or xe2x80x94N(H)OH;
each R8 is independently -alkyl, -cycloalkyl, -aryl, -heteroaryl, -heterocyclyl, -alkylcycloalkyl -alkylaryl, -alkylheteroaryl, or -alkylheterocyclyl, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11, and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1;
each R9 is independently -aryl, -heteroaryl, cycloalkyl, or -heterocyclyl, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11, and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1;
each R10 is independently xe2x80x94OH, xe2x80x94SH, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94CO2H, xe2x80x94C(O)NH2, xe2x80x94N(H)C(O)H, xe2x80x94N(H)C(O)NH2, -perfluoroalkyl, xe2x80x94O-alkyl, xe2x80x94O-aryl, xe2x80x94O-alkylaryl, xe2x80x94N(H)alkyl, xe2x80x94N(H)aryl, xe2x80x94N(H)-alkylaryl, xe2x80x94N(alkyl)2, xe2x80x94C(O)N(H)alkyl, xe2x80x94C(O)N(alkyl)2, xe2x80x94N(H)C(O)alkyl, xe2x80x94N(H)C(O)N(H)alkyl, xe2x80x94N(H)C(O)N(alkyl)2, xe2x80x94S-alkyl, xe2x80x94S-aryl, xe2x80x94S-alkylaryl, xe2x80x94S(O)2alkyl, xe2x80x94S(O)alkyl, xe2x80x94C(O)alkyl, xe2x80x94CH2NH2, xe2x80x94CH2N(H)alkyl, or xe2x80x94CH2N(alkyl)2, -alkyl, -cycloalkyl, -aryl, -heteroaryl, -heterocyclyl, -alkylcycloalkyl -alkylaryl, -alkylheteroaryl, or -alkylheterocyclyl, wherein a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11 and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1; and
each R11 is independently xe2x80x94OH, xe2x80x94SH, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94CO2H, xe2x80x94C(O)NH2, xe2x80x94N(H)C(O)H, xe2x80x94N(H)C(O)NH2, -alkyl, -cycloalkyl, -perfluoroalkyl, xe2x80x94O-alkyl, xe2x80x94O-aryl, xe2x80x94O-alkylaryl, xe2x80x94N(H)alkyl, xe2x80x94N(H)aryl, xe2x80x94N(H)-alkylaryl, xe2x80x94N(alkyl)2, xe2x80x94C(O)N(H)alkyl, xe2x80x94C(O)N(alkyl)2, xe2x80x94N(H)C (O)alkyl, xe2x80x94N(H)C()N(H)alkyl, xe2x80x94N(H)C(O)N(alkyl)2, xe2x80x94S-alkyl, xe2x80x94S-aryl, xe2x80x94S-alkylaryl, xe2x80x94S(O)2alkyl, xe2x80x94S(O)alkyl, xe2x80x94C(O)alkyl, xe2x80x94CH2NH2, xe2x80x94CH2N(H)alkyl, or xe2x80x94CH2N(alkyl)2.
In an alternative form of embodiment A:
R1 is H, xe2x80x94R8, xe2x80x94C(O)R8, xe2x80x94C(O)C(O)R8, xe2x80x94S(O)2R8, xe2x80x94S(O)R8, xe2x80x94C(O)OR8, xe2x80x94C(O)N(H)R8, xe2x80x94S(O)2N(H)xe2x80x94R8, xe2x80x94S(O)N(H)xe2x80x94R8, xe2x80x94C(O)C(O)N(H)R8, xe2x80x94C(O)CHxe2x95x90CHR8, xe2x80x94C(O)CH2OR8, xe2x80x94C(O)CH2N(H)R8, xe2x80x94C(O)N(R8)2, xe2x80x94S(O)2N(R8)2, xe2x80x94S(O)N(R8)2, xe2x80x94C(O)C(O)N(R8)2, xe2x80x94C(O)CH2N(R8)2, CH2R8, xe2x80x94CH2-alkenyl-R8, or xe2x80x94CH2-alkynyl-R8;
R2 is xe2x80x94H and each R3 is independently xe2x80x94H, an amino acid side chain, xe2x80x94R8, alkenyl-R9, or alkynyl-R9, or each R3, together with the atom to which they are bound, form a 3 to 7 membered cyclic or heterocyclic cyclic ring system, or R2 and one R3 together with the atoms to which they are bound, form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by xe2x80x94R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by xe2x80x94R11, a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced by xe2x80x94R1;
each R10 is independently xe2x80x94OH, xe2x80x94SH, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94CO2H, xe2x80x94C(O)NH2, xe2x80x94N(H)C(O)H, xe2x80x94N(H)C(O)NH2, -perfluoroalkyl, xe2x80x94O-alkyl, xe2x80x94O-aryl, xe2x80x94O-alkylaryl, xe2x80x94N(H)alkyl, xe2x80x94N(H)aryl, xe2x80x94N(H)-alkylaryl, xe2x80x94N(alkyl)2, xe2x80x94C(O)N(H)alkyl, xe2x80x94C(O)N(alkyl)2, xe2x80x94N(H)C(O)alkyl, xe2x80x94N(H)C(O)Oalkyl, xe2x80x94N(H)C(O)Oaryl, xe2x80x94N(H)C(O)Oalkylaryl, xe2x80x94N(H)C(O)Oheteroaryl, xe2x80x94N(H)C(O)Oalkylheteroaryl, xe2x80x94N(H)C(O)Ocycloalkyl, xe2x80x94N(H)C(O)N(H)alkyl, xe2x80x94N(H)C(O)N(alkyl)2, xe2x80x94N(H)C(O)N(H)aryl, xe2x80x94N(H)C(O)N(H)alkylaryl, xe2x80x94N(H)C(O)N(H)heteroaryl, xe2x80x94N(H)C(O)N(H)alkylheteroaryl, xe2x80x94N(H)C(O)N(H)cycloalkyl, xe2x80x94S-alkyl, xe2x80x94S-aryl, xe2x80x94S-alkylaryl, xe2x80x94S(O)2alkyl, xe2x80x94S(O)alkyl, xe2x80x94C(O)alkyl, xe2x80x94CH2NH2, xe2x80x94CH2N(H)alkyl, or xe2x80x94CH2N(alkyl)2, -alkyl, -cycloalkyl, -aryl, -heteroaryl, -heterocyclyl, -alkylcycloalkyl -alkylaryl, -alkylheteroaryl, or -alkylheterocyclyl, wherein a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11 and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1; and
the other substituents are as defined above.
Preferably, in any of the above embodiments:
m is 0;
R2 is xe2x80x94H;
one R3 is xe2x80x94H and the other R3 is xe2x80x94R8, -alkenyl-R9, or -alkynyl-R9; or
R4 and one R5 together with the atoms to which they are bound form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11, and a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced with R1, wherein the ring system is: 
In an alternative preferred embodiment, X is C(R3)2 or one R3 is an amino acid side chain, xe2x80x94R8, alkenyl-R9, or alkynyl-R9.
More preferably, one R3 is xe2x80x94H and the other R3 is -alkyl; or
R4 and one R5 together with the atoms to which they are bound form a 3 to 7 membered cyclic or heterocyclic ring system, wherein any hydrogen atom bound to a carbon atom of the ring system is optionally replaced by R10 and any hydrogen atom bound to a nitrogen atom of the ring system is optionally replaced by R1, selected from: 
Most preferably, one R3 is xe2x80x94H and the other R3 is xe2x80x94C(H)(CH3)2 or xe2x80x94C(CH3)3; and
R4 and one R5 together with the atoms to which they are bound form a 3 to 7 membered cyclic or heterocyclic ring system, wherein any hydrogen atom bound to a carbon atom of the ring system is optionally replaced by R10 and any hydrogen atom bound to a nitrogen atom of the ring system is optionally replaced by R1, selected from: 
In an alternative most preferred embodiment, one R3 is xe2x80x94H and the other R3 is xe2x80x94CH3, xe2x80x94C(H)(CH3)2 or xe2x80x94C(CH3)3 and R4 and R5 are as defined directly above.
According to another embodiment B, the present invention provides a compound of formula I, wherein Y is: 
provided that when R6 is not hydrogen, R6 and Y, together with the nitrogen to which they are bound, form a ring (g): 
R12 is xe2x80x94C(O)alkyl, xe2x80x94C(O)cycloalkyl, xe2x80x94C(O)alkyenyl, xe2x80x94C(O)alkylaryl, xe2x80x94C(O)alkylheteroaryl, xe2x80x94C(O) heterocyclyl, or xe2x80x94C(O)alkylheterocyclyl;
R13 is xe2x80x94H, -alkyl, -aryl, -alkylaryl or -alkylheteroaryl; and
the other substituents are as described above.
Preferably, in (c), (d), (e), or (f), R8 is methyl, ethyl, n-propyl, isopropyl, cyclopentyl, phenethyl, or benzyl.
Preferred definitions for the other individual components of embodiment B are the same as those set forth above for embodiment A.
A preferred embodiment C of this invention provides compounds of formula I: 
wherein:
Y is: 
m is 0 or 1;
X is xe2x80x94C(R3)2xe2x80x94
R1 is H, xe2x80x94R8, xe2x80x94C(O)R8, xe2x80x94C(O)C(O)R8, xe2x80x94S(O)2R8, xe2x80x94S(O)R8, xe2x80x94C(O)OR8, xe2x80x94C(O)N(H)R8, xe2x80x94S(O)2N(H)xe2x80x94R8, xe2x80x94S(O)N(H)xe2x80x94R8, xe2x80x94C(O)C(O)N(H)R8, xe2x80x94C(O)CHxe2x95x90CHR8, xe2x80x94C(O)CH2OR8, xe2x80x94C(O)CH2N(H)R8, xe2x80x94C(O)N(R8)2, xe2x80x94S(O)2N(R8)2, xe2x80x94S(O)N(R8)2, xe2x80x94C(O)C(O)N(R8)2, xe2x80x94C(O)CH2N(R8)2, xe2x80x94CH2R8, xe2x80x94CH2-alkenyl-R8, or xe2x80x94CH2-alkynyl-R8;
R2 is xe2x80x94H and each R3 is independently xe2x80x94H, an amino acid side chain, xe2x80x94R8, alkenyl-R9, or alkynyl-R9, or each R3 together with the atom to which they are bound, form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by xe2x80x94R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by xe2x80x94R11, a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced by xe2x80x94R1;
R4 is xe2x80x94H and each R5 is independently xe2x80x94H, an amino acid side chain, xe2x80x94R8, -alkenyl-R9, or -alkynyl-R9, or R4 and one R5 together with the atoms to which they are bound form a 3 to 7 membered cyclic or heterocyclic ring system, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11, and a hydrogen atom bound to any nitrogen atom of the ring system is optionally replaced with R1;
R6 is xe2x80x94H;
R7 is xe2x80x94OH, xe2x80x94OR8, xe2x80x94N(H)OH, or xe2x80x94N(H)S(O)2R8;
each R8 is independently -alkyl, -cycloalkyl, -aryl, -heteroaryl, -heterocyclyl, -alkylcycloalkyl -alkylaryl, -alkylheteroaryl, or -alkylheterocyclyl, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11 and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1;
each R9 is independently -aryl, -heteroaryl, cycloalkyl, or -heterocyclyl, wherein a hydrogen atom bound to any -alkyl or -cycloalkyl carbon atom is optionally replaced by R10, a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11, and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1;
each R10 is independently xe2x80x94OH, xe2x80x94SH, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94CO2H, xe2x80x94C(O)NH2, xe2x80x94N(H)C(O)H, xe2x80x94N(H)C(O)NH2, -perfluoroalkyl, xe2x80x94O-alkyl, xe2x80x94O-aryl, xe2x80x94O-alkylaryl, xe2x80x94N(H)alkyl, xe2x80x94N(H)aryl, xe2x80x94N(H)-alkylaryl, xe2x80x94N(alkyl)2, xe2x80x94C(O)N(H)alkyl, xe2x80x94C(O)N(alkyl)2, xe2x80x94N(H)C(O)alkyl, xe2x80x94N(H)C(O)Oalkyl, xe2x80x94N(H)C(O)Oaryl, xe2x80x94N(H)C(O)Oalkylaryl, xe2x80x94N(H)C(O)Oheteroaryl, xe2x80x94N(H)C(O)Oalkylheteroaryl, xe2x80x94N(H)C(O)Ocycloalkyl, xe2x80x94N(H)C(O)N(H)alkyl, xe2x80x94N(H)C(O)N(alkyl)2, xe2x80x94N(H)C(O)N(H)aryl, xe2x80x94N(H)C(O)N(H)alkylaryl, xe2x80x94N(H)C(O)N(H)heteroaryl, xe2x80x94N(H)C(O)N(H)alkylheteroaryl, xe2x80x94N(H)C(O)N(H)cycloalkyl, xe2x80x94S-alkyl, xe2x80x94S-aryl, xe2x80x94S-alkylaryl, xe2x80x94S(O)2alkyl, xe2x80x94S(O)alkyl, xe2x80x94C(O)alkyl, xe2x80x94CH2NH2, xe2x80x94CH2N(H)alkyl, or xe2x80x94CH2N(alkyl)2, -alkyl, -cycloalkyl, -aryl, -heteroaryl, -heterocyclyl, -alkylcycloalkyl -alkylaryl, -alkylheteroaryl, or -alkylheterocyclyl, wherein a hydrogen atom bound to any -aryl or -heteroaryl carbon atom is optionally replaced by R11 and a hydrogen atom bound to any nitrogen atom is optionally replaced by R1; and
each R11 is independently xe2x80x94OH, xe2x80x94SH, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, xe2x80x94I, xe2x80x94NO2, xe2x80x94CN, xe2x80x94NH2, xe2x80x94CO2H, xe2x80x94C(O)NH2, xe2x80x94N(H)C(O)H, xe2x80x94N(H)C(O)NH2, -alkyl, -cycloalkyl, -perfluoroalkyl, xe2x80x94O-alkyl-, xe2x80x94O-aryl, xe2x80x94O-alkylaryl, xe2x80x94N(H)alkyl, xe2x80x94N(H)aryl, xe2x80x94N(H)-alkylaryl, xe2x80x94N(alkyl)2, xe2x80x94C(O)N(H)alkyl, xe2x80x94C(O)N(alkyl)2, xe2x80x94N(H)C(O)alkyl, xe2x80x94N(H)C(O)N(H)alkyl, xe2x80x94N(H)C(O)N(alkyl)2, xe2x80x94S-alkyl, xe2x80x94S-aryl, xe2x80x94S-alkylaryl, xe2x80x94S(O)2alkyl, xe2x80x94S(O)alkyl, xe2x80x94C(O)alkyl, xe2x80x94CH2NH2, xe2x80x94CH2N(H)alkyl, or xe2x80x94CH2N(alkyl)2;
provided that if one R3 is xe2x80x94H, then the other R3 is not xe2x80x94H.
Another preferred embodiment D of the present invention provides a compound of formula I, wherein Y is: 
R12 is xe2x80x94C(O)alkyl, xe2x80x94C(O)cycloalkyl, xe2x80x94C(O)alkyenyl, xe2x80x94C(O)alkylaryl, xe2x80x94C(O)alkylheteroaryl, xe2x80x94C(O)heterocyclyl, or xe2x80x94C(O)alkylheterocyclyl; and
the other substituents are as described above except that both of the R3 groups may be xe2x80x94H.
In any of embodiments A-D, preferred compounds are those whererin:
R1 is xe2x80x94C(O)R8 or xe2x80x94C(O)(O)R8;
R2 and one R3 are both xe2x80x94H and the other R3 is an amino acid side chain, xe2x80x94R8, alkenyl-R9, or alkynyl-R9; or
R4 and one R5 together with the atoms to which they are bound form a ring system selected from: 
provided that each of the ring systems are optionally substituted with one or more R10 groups.
Alternatively, preferred compounds of embodiments A-D are those wherein R3 is xe2x80x94H and the other R3 is methyl, isopropyl, tert-butyl, xe2x80x94CH2SR8, xe2x80x94CH2SO2R8, xe2x80x94CH2CH2SR8, xe2x80x94CH2CH2SO2R8,
More preferred compounds of embodiments A-D are those wherein R4 and one R5 together with the atoms to which they are bound form the ring system and: 
the other R5 is H; or
one R3 is xe2x80x94H and the other R3 is methyl.
Alternatively, more preferred compounds of embodiments A-D are those wherein R4 and one R5 together with the atoms to which they are bound form the ring system: 
and the other R5 is H.
In the above alternative embodiment, R10 is preferably, 4-fluoro or 4,4-difluoro.
Most preferred compounds of this invention are those wherein R3 is methyl; and
R4 and one R5 together with the atoms to which they are bound form the ring system: 
and the other R5 is H.
Alternatively, most preferred compounds of embodiments A-D are those wherein R3 is methyl; and R4 and one R5 together with the atoms to which they are bound form the ring system: and 
the other R5 is H; and
R10 is 4-fluoro or 4,4-difluoro.
Preferred compounds of embodiments (B) or (D) are those wherein Y is: 
wherein Z represents xe2x80x94OR and Z is: CH3O, 
Specific compounds of this invention include, but are not limited to, Examples 5a-5bd, 7a-7at, 9a-9g, 15a-15f, 16a-16b, 17a-17e, 18a-18f, 20a-20t, 23a-23i, 24a-24e, 25a-25e, 26a-26h, 27a-27n, 28a-28c, 29a-29s, 32a-32e, 34, G1, G2, 41, 42, 45, 46, 51, 52, 56, 57, 60, 61, 64, 65, 68, 69, 72, 73, 76-93, 98a-z, aa-az, and ba-bb, 101, 102a, 102b, 108a-d, 110, 111, 116a-h, 120a and b, 121, 122a-v, and 123a-c.
The compounds of this invention may contain one or more xe2x80x9casymmetricxe2x80x9d carbon atoms and thus may occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. Each stereogenic carbon may be of the R or S configuration. Although specific compounds and scaffolds exemplified in this application may be depicted in a particular stereochemical configuration, compounds and scaffolds having either the opposite stereochemistry at any given chiral center or mixtures thereof are also envisioned.
All such isomeric forms of these compounds are expressly included in the present invention, as well as pharmaceutically acceptable derivative thereof.
The term xe2x80x9cpharmaceutically acceptable derivativexe2x80x9d means any pharmaceutically acceptable salt, ester, or salt of such ester, of a compound of this invention or any other compound which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of this invention or an active metabolite or residue thereof.
Pharmaceutically acceptable salts of the compounds of this invention include, for example, those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene-p-sulfonic, tartaric, acetic, citric, methanesulfonic, formic, benzoic, malonic, naphthalene-2-sulfonic and benzenesulfonic acids. Other acids, such as oxalic, while not in themselves pharmaceutically acceptable, may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts. Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and Nxe2x80x94(C1-4 alkyl)4+salts.
This invention also envisions the xe2x80x9cquaternizationxe2x80x9d of any basic nitrogen-containing groups of the compounds disclosed herein. The basic nitrogen can be quaternized with any agents known to those of ordinary skill in the art including, for example, lower alkyl halides, such as methyl, ethyl, propyl and butyl chloride, bromides and iodides; dialkyl sulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; and aralkyl halides including benzyl and phenethyl bromides. Water or oil-soluble or dispersible products may be obtained by such quaternization.
When multiply substituted, each substituent may be picked independently of any other substituent as long as the combination of substituents results in the formation of a stable compound.
Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term xe2x80x9cstablexe2x80x9d, as used herein, refers to compounds which possess stability sufficient to allow manufacture and administration to a mammal by methods known in the art. Typically, such compounds are stable at a temperature of 40xc2x0 C. or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
Preferred compounds of this invention may be readily absorbed by the bloodstream of patients upon oral administration. This oral availability makes such compounds excellent agents for orally-administered treatment and prevention regimens against IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated diseases.
It should be understood that the compounds of this invention may exist in various equilibrium forms, depending on conditions including choice of solvent, pH, and others known to the practitioner skilled in the art. All such forms of these compounds are expressly included in the present invention. In particular, many of the compounds of this invention, especially those which contain aldehyde or ketone groups and carboxylic acid groups in Y, may take hemi-acetal or hydrated forms. For example, compounds of embodiment A are in a hemiacetal form when Y is: 
Depending on the choice of solvent and other conditions known to the practitioner skilled in the art, compounds of this invention may also take hydrated, acyloxy acetal, acetal, or enol forms. For example, compounds of this invention are in hydrated forms when Y is: 
and R8 is H;
acyloxy acetal forms when Y is: 
acetal forms when Y is and R8 is other than H: 
and enol forms when Y is: 
In addition, it should be understood that the equilibrium forms of the compounds of this invention may include tautomeric forms. All such forms of these compounds are expressly included in the present invention.
The compounds of formula I may be synthesized using conventional techniques. Advantageously, these compounds are conveniently synthesized from readily available starting materials.
Compounds of this invention may be prepared using the processes described herein. As can be appreciated by the skilled practitioner, these processes are not the only means by which the compounds described and claimed in this application may be synthesized. Further methods will be evident to those of ordinary skill in the art. Additionally, the various synthetic steps described herein may be performed in an alternate sequence or order to give the desired compounds.
It should be understood that the compounds of this invention may be modified by appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion. In addition, the compounds may be altered to pro-drug form such that the desired compound is created in the body of the patient as the result of the action of metabolic or other biochemical processes on the pro-drug. Such pro-drug forms typically demonstrate little or no activity in in vitro assays. Some examples of pro-drug forms include ketal, acetal, oxime, imine and hydrazone forms of compounds which contain ketone or aldehyde groups, especially where they occur in the Y group of the compounds of this invention. Other examples of pro-drug forms include the hemi-ketal, hemi-acetal, acyloxy ketal, acyloxy acetal, ketal, acetal and enol forms that are described herein.
The compounds of this invention are caspase inhibitors, and in particular ICE inhibitors. Accordingly, these compounds are capable of targeting and inhibiting events in IL-1-, apoptosis-, IGIF-, and IFN-xcex3-mediated diseases, and, thus, the ultimate activity of that protein in inflammatory diseases, autoimmune diseases, destructive bone, proliferative disorders, infectious diseases, and degenerative diseases. For example, the compounds of this invention inhibit the conversion of precursor IL-1xcex2 to mature IL-1xcex2 by inhibiting ICE. Because ICE is essential for the production of mature IL-1, inhibition of that enzyme effectively blocks initiation of IL-1-mediated physiological effects and symptoms, such as inflammation, by inhibiting the production of mature IL-1. Thus, by inhibiting IL-1xcex2 precursor activity, the compounds of this invention effectively function as IL-1 inhibitors.
Compounds of this invention also inhibit conversion of pro-IGIF into active, mature IGIF by inhibiting ICE. Because ICE is essential for the production of mature IGIF, inhibition of ICE effectively blocks initiation of IGIF-mediated physiological effects and symptoms, by inhibiting production of mature IGIF. IGIF is in turn essential for the production of IFN-xcex3. ICE therefore effectively blocks initiation of IFN-xcex3-mediated physiological effects and symptoms, by inhibiting production of mature IGIF and thus production of IFN-xcex3.
The compounds of this invention are surprisingly bioavailable when compared with peptidyl inhibitors, such as those described in, for example, EP 618 223, EP 623 592, WO 93/09135, WO 93/16710, U.S. Pat. No. 5,434,248, WO 95/35308, or WO 96/33209. Thus, the pharmaceutical compositions and methods of this invention will be useful for controlling caspase activity in vivo. The compositions and methods of this invention will therefore be useful for controlling IL-1, IGIF, or IFN-xcex3 levels in vivo and for treating or reducing the advancement, severity or effects of IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated conditions, including diseases, disorders or effects.
Pharmaceutical compositions of this invention comprise a compound of formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier. Such compositions may optionally comprise an additional therapeutic agent. Such agents include, but are not limited to, an anti-inflammatory agent, a matrix metalloprotease inhibitor, a lipoxygenase inhibitor, a cytokine antagonist, an immunosuppressant, an anti-cancer agent, an anti-viral agent, a cytokine, a growth factor, an immunomodulator, a prostaglandin or an anti-vascular hyperproliferation compound.
The term xe2x80x9cpharmaceutically acceptable carrierxe2x80x9d refers to a non-toxic carrier that may be administered to a patient, together with a compound of this invention, and which does not destroy the pharmacological activity thereof.
Pharmaceutically acceptable carriers that may be used in the pharmaceutical compositions of this invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, wool fat and self-emulsifying drug delivery systems (SEDDS) such as a-tocopherol, polyethyleneglycol 1000 succinate, or other similar polymeric delivery matrices.
In pharmaceutical composition comprising only a compound of embodiments A-D as the active component, methods for administering these compositions may additionally comprise the step of administering to the subject an additional agent. Such agents include, but are not limited to, an anti-inflammatory agent, a matrix metalloprotease inhibitor, a lipoxygenase inhibitor, a cytokine antagonist, an immunosuppressant, an anti-cancer agent, an anti-viral agent, a cytokine, a growth factor, an immunomodulator, a prostaglandin or an anti-vascular hyperproliferation compound.
The term xe2x80x9cpharmaceutically effective amountxe2x80x9d refers to an amount effective in treating or ameliorating an IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated disease in a patient. The term xe2x80x9cprophylactically effective amountxe2x80x9d refers to an amount effective in preventing or substantially lessening IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated diseases in a patient.
The compounds of this invention may be employed in a conventional manner for controlling IGIF and IFN-xcex3 levels in vivo and for treating diseases or reducing the advancement or severity of effects which are mediated by IL-1, apoptosis, IGIF, or IFN-xcex3. Such methods of treatment, their dosage levels and requirements may be selected by those of ordinary skill in the art from available methods and techniques.
For example, a compound of this invention may be combined with a pharmaceutically acceptable adjuvant for administration to a patient suffering from an IL-1-, apoptosis-, IGIF-, or IFN-xcex3-mediated disease in a pharmaceutically acceptable manner and in an amount effective to lessen the severity of that disease.
Alternatively, the compounds of this invention may be used in compositions and methods for treating or protecting individuals against IL-1, apoptosis-, IGIF, or IFN-xcex3 mediated diseases over extended periods of time. The compounds may be employed in such compositions either alone or together with other compounds of this invention in a manner consistent with the conventional utilization of enzyme. inhibitors in pharmaceutical compositions. For example, a compound of this invention may be combined with pharmaceutically acceptable adjuvants conventionally employed in vaccines and administered in prophylactically effective amounts to protect individuals over an extended period of time against IL-1-, apoptosis-, IGIF, or IFN-xcex3 mediated diseases.
The compounds of formula I may also be co-administered with other caspase or ICE inhibitors to increase the effect of therapy or prophylaxis against various IL-1-, apoptosis-, IGIF-, or IFN-xcex3 mediated diseases.
In addition, the compounds of this invention may be used in combination either conventional anti-inflammatory agents or with matrix metalloprotease inhibitors, lipoxygenase inhibitors and antagonists of cytokines other than IL-1xcex2.
The compounds of this invention can also be administered in combination with immunomodulators (e.g., bropirimine, anti-human alpha-interferon antibody, IL-2, GM-CSF, methionine enkephalin, interferon-alpha, diethyldithiocarbamate, tumor necrosis factor, naltrexone and EPO), with prostaglandins, or with antiviral agents (e.g., 3TC, polysulfated polysaccharides, ganiclovir, ribavirin, acyclovir, alpha interferon, trimethotrexate and fancyclovir) or prodrugs of these or related compounds to prevent or combat IL-1-mediated disease symptoms such as inflammation.
When the compounds of this invention are administered in combination therapies with other agents, they may be administered sequentially or concurrently to the patient. Alternatively, pharmaceutical or prophylactic compositions according to this invention comprise a combination of a compound of formula I and another therapeutic or prophylactic agent.
The pharmaceutical compositions of this invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. We prefer oral administration. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically-acceptable carriers, adjuvants or vehicles. In some cases, the pH of the formulation may be adjusted with pharmaceutically acceptable acids, bases or buffers to enhance the stability of the formulated compound or its delivery form. The term parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are mannitol, water, Ringer""s solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil, especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as those described in Pharmacopeia Helvetica, or a similar alcohol.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspensions and solutions. In the case of tablets for oral use, carriers which are commonly used include lactose and corn starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions and solutions and propylene glycol are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
The pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax and polyethylene glycols.
Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical composition can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tract by rectal suppository formulation or in a suitable enema formulation. Topically-administered transdermal patches are also included in this invention.
The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
Dosage levels of between about 0.01 and about 100 mg/kg body weight per day, preferably between 0.5 and about 75 mg/kg body weight per day and most preferably between about 1 and 50 mg/kg body weight per day of the active ingredient compound are useful in a monotherapy for the prevention and treatment of IL-1-, apoptosis-, IGIF-, and IFN-xcex3 mediated diseases, including inflammatory diseases, autoimmune diseases, destructive bone disorders, proliferative disorders, infectious diseases, degenerative diseases, necrotic diseases, inflammatory peritonitis, osteoarthritis, acute pancreatitis, chronic pancreatitis, asthma, adult respiratory distress syndrome, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves"" disease, autoimmune gastritis, insulin-dependent diabetes mellitus (Type I), autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, chronic active hepatitis, myasthenia gravis, inflammatory bowel disease, Crohn""s disease, psoriasis, atopic dermatitis, graft vs. host disease, osteoporosis, multiple myeloma-related bone disorder, leukemias and related disorders, myelodysplastic syndrome, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi""s sarcoma, multiple myeloma, sepsis, septic shock, Shigellosis, Alzheimer""s disease, Parkinson""s disease, cerebral ischemia, myocardial ischemia, spinal muscular atrophy, multiple sclerosis, AIDS-related encephalitis, HIV-related encephalitis, aging, alopecia, neurological damage due to stroke, ulcerative collitis, infectious hepatitis, juvenile diabetes, lichenplanus, acute dermatomyositis, eczema, primary cirrhosis, uveitis, Behcet""s disease, atopic skin disease, pure red cell aplasia, aplastic anemia, amyotrophic lateral sclerosis, nephrotic syndrome and systemic diseases or diseases with effects localized in the liver or other organs having an inflammatory or apoptotic component caused by excess dietary alcohol intake or viruses, such as HBV, HCV, HGV, yellow fever virus, dengue fever virus, and Japanese encephalitis virus.
Typically, the pharmaceutical compositions of this invention will be administered from about 1 to 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the host treated and the particular mode of administration. A typical preparation will contain from about 5% to about 95% active compound (w/w). Preferably, such preparations contain from about 20% to about 80% active compound.
When the compositions of this invention comprise a combination of a compound of formula I and one or more additional therapeutic or prophylactic agents, both the compound and the additional agent should be present at dosage levels of between about 10% to 80% of the dosage normally administered in a monotherapy regime.
Upon improvement of a patient""s condition, a maintenance dose of a compound, composition or combination of this invention may be administered, if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained when the symptoms have been alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis upon any recurrence or disease symptoms.
As the skilled artisan will appreciate, lower or higher doses than those recited above may be required. Specific dosage and treatment regimens for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combination, the severity and course of the disease, and the patient""s disposition to the disease and the judgment of the treating physician.
IL-1 or apoptosis mediated diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, inflammatory diseases-, autoimmune diseases, proliferative disorders, infectious diseases, and degenerative diseases. The apoptosis-mediated diseases which may be treated or prevented by the compounds of this invention include degenerative diseases.
IL-1 or apoptosis mediated inflammatory diseases which may be treated or prevented include, but are not limited to osteoarthritis, acute pancreatitis, chronic pancreatitis, asthma, and adult respiratory distress syndrome. Preferably the inflammatory disease is osteoarthritis or acute pancreatitis.
IL-1 or apoptosis mediated autoimmune diseases which may be treated or prevented include, but are not limited to, glomerulonephritis, rheumatoid arthritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves"" disease, autoimmune gastritis, insulin-dependent diabetes mellitus (Type I), autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, chronic active hepatitis, myasthenia gravis, multiple sclerosis, inflammatory bowel disease, Crohn""s disease, psoriasis, atopic dermatitis and graft vs. host disease. Preferably the autoimmune disease is rheumatoid arthritis, inflammatory bowel disease, Crohn""s disease, psoriasis, or atopic dermatitis.
IL-1 or apoptosis mediated destructive bone disorders which may be treated or prevented include, but are not limited to, osteoporosis and multiple myeloma-related bone disorder.
IL-1 or apoptosis mediated proliferative diseases which may be treated or prevented include, but are not limited to, leukemias and related disorders, such as myelodysplastic syndrome, acute myelogenous leukemia, chronic myelogenous leukemia, metastatic melanoma, Kaposi""s sarcoma, and multiple myeloma.
IL-1 or apoptosis mediated infectious diseases which may be treated or prevented include, but are not limited to, sepsis, septic shock, and Shigellosis.
IL-1 or apoptosis mediated degenerative or necrotic diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, Alzheimer""s disease, Parkinson""s disease, cerebral ischemia, and myocardial ischemia. Preferably, the degenerative disease is Alzheimer""s disease.
IL-1 or apoptosis-mediated degenerative diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, Alzheimer""s disease, Parkinson""s disease, cerebral ischemia, myocardial ischemia, spinal muscular atrophy, multiple sclerosis, AIDS-related encephalitis, HIV-related encephalitis, aging, alopecia, and neurological damage due to stroke.
Other diseases having an inflammatory or apoptotic component may be treated or prevented by the compounds of this invention. Such diseases may be systemic diseases or diseases with effects localized in the liver or other organs and may be caused by, for example, excess dietary alcohol intake or viruses, such as HBV, HCV, HGV, yellow fever virus, dengue fever virus, and Japanese encephalitis virus.
IGIF- or IFN-xcex3-mediated diseases which may be treated or prevented by the compounds of this invention include, but are not limited to, inflammatory, infectious, autoimmune, proliferative, neurodegenerative and necrotic conditions.
IGIF- or IFN-xcex3-mediated inflammatory diseases which may be treated or prevented include, but are not limited to osteoarthritis, acute pancreatitis, chronic pancreatitis, asthma, rheumatoid arthritis, inflammatory bowel disease, Crohn""s disease, ulcerative collitis, cerebral ischemia, myocardial ischemia and adult respiratory distress syndrome. Preferably, the inflammatory disease is rheumatoid arthritis, ulcerative collitis, Crohn""s disease, hepatitis or adult respiratory distress syndrome.
IGIF- or IFN-xcex3-mediated infectious diseases which may be treated or prevented include, but are not limited to infectious hepatitis, sepsis, septic shock and Shigellosis.
IGIF- or IFN-xcex3-mediated autoimmune diseases which may be treated or prevented include, but are not limited to glomerulonephritis, systemic lupus erythematosus, scleroderma, chronic thyroiditis, Graves"" disease, autoimmune gastritis, insulin-dependent diabetes mellitus (Type I), juvenile diabetes, autoimmune hemolytic anemia, autoimmune neutropenia, thrombocytopenia, myasthenia gravis, multiple sclerosis, psoriasis, lichenplanus, graft vs. host disease, acute dermatomyositis, eczema, primary cirrhosis, hepatitis, uveitis, Behcet""s disease, atopic skin disease, pure red cell aplasia, aplastic anemia, amyotrophic lateral sclerosis and nephrotic syndrome. Preferably, the autoimmune disease is glomerulonephritis, insulin-dependent diabetes mellitus (Type I), juvenile diabetes, psoriasis, graft vs. host disease or hepatitis.
More preferred diseases which may be treated or prevented include rheumatoid arthritis, inflammatory bowel disease, including Crohn""s disease and ulcerative colitis, inflammatory peritonitis, septic shock, pancreatitis, traumatic brain injury, organ transplant rejection, osteoarthritis, asthma, psoriasis, Alzeheimer""s disease, atopic dermatitis, or leukemias and related disorders, such as myelodysplastic syndrome or multiple myeloma.
Accordingly, one embodiment of this invention provides a method for treating or preventing an IL-1 or apoptosis mediated disease in a subject comprising the step of administering to the subject any compound, pharmaceutical composition, or combination described herein and a pharmaceutically acceptable carrier.
Another embodiment of this invention provides a method for decreasing IGIF production in a subject comprising the step of administering to the subject any compound, pharmaceutical composition, or combination described herein and a pharmaceutically acceptable carrier.
Yet another embodiment of this invention provides a method for decreasing IFN-xcex3 production in a subject comprising the step of administering to the subject any compound, pharmaceutical composition, or combination described herein and a pharmaceutically acceptable carrier.
Although this invention focuses on the use of the compounds disclosed herein for preventing and treating IL-1, apoptosis-, IGIF, and IFN-xcex3-mediated diseases, the compounds of this invention can also be used as inhibitory agents for other cysteine proteases.
The compounds of this invention are also useful as commercial reagents which effectively bind to caspases or other cysteine proteases including, but not limited to ICE. As commercial reagents, the compounds of this invention, and their derivatives, may be used to block proteolysis of a target peptide in biochemical or cellular assays for ICE and ICE homologs or may be derivatized to bind to a stable resin as a tethered substrate for affinity chromatography applications. These and other uses which characterize commercial cysteine protease inhibitors will be evident to those of ordinary skill in the art.
In order that this invention be more fully understood, the following examples are set forth. These examples are for the purpose of illustration only and are not to be construed as limiting the scope of the invention in any way.
Analytical HPLC Conditions:
Column: C-18, Particle size: 5xcexc, Pore size: 100 xc3x85,
Column size: 4.6xc3x97150 mm
Solvent A: 0.1% TFA/1% MeCN/98.9% water
Solvent B: 0.1% TFA/99.9% MeCN
Gradient: A to B over 20 min at a flow rate of 1 mL/min
Column: Cyano, Particle size: 5xcexc, Pore size: 100 xc3x85,
Column size: 4.6xc3x97150 mm
Solvent A: 0.1% TFA/1% MeCN/98.9% water
Solvent B: 0.1% TFA/99.9% MeCN
Gradient: A/B=99%/1% to 50%/50% over 20 min at a flow rate of 1 mL/min
HPLC Mass Spectral Analysis
Mass Spectral Analysis: All mass spectral data were collected using a Micromass Quattro II triple quadrupole mass spectrometer (Beverly, Mass.) equipped with a cross-flow electrospray ionization source. The mass spectrometer was coupled to a HPLC system manufactured by Hewlett-Packard (HP1100). The autosampler for the system was a Gilson 215 (Middleton, Wis.) liquid handler. All of the equipment was controlled by the MassLynx software package purchased from Micromass.
Mass spectral analysis was performed by liquid chromatography-MS to determine purity and confirm molecular weight simultaneously. In instances where the sample purity had been determined by other means, a flow injection analysis (FIA) was used instead of the full chromatography analysis. In all cases, both positive and negative ion spectra were collected.
Mass Spectrum Acquisition Conditions: For all experiments, the mass spectrometer was configured in electrospray mode with the cross-flow counter electrode. A flow splitter was used to reduce the flow from the HPLC to 40% of the original flow. The inlet temperature was set to 140xc2x0 C. and the drying gas flow was set to maximize signal. The resolution of the mass spectrometer was adjusted to 0.65 amu FWHM and data was collected in centroid mode. In positive ion mode, the cone voltage was set to 25V, the capillary voltage was 3.8 kV. In negative ion mode, the cone voltage was set to 25 V and the capillary voltage was set to 3.5 kV. In both positive and negative ion mode, the time to acquire a full spectrum was is with a switching time of 0.25 seconds between scans. The mass range scanned for molecules with an expected molecular weight of less than 350 amu was 70-500 m/z while for molecules with a expected mass of more than 350 amu the mass to charge ratio scanned was 200-1000 m/z.
Chromatography Conditions: Liquid chromatography was performed using a YMC AQ C18 column (150 mmxc3x973 mm with S1 m particle and a 120 xc3x85 pore size). For all analysis, MeCN with 0.2% formic acid was combined with water with 0.2% formic acid to form the elution gradient. The gradient profile consisted of starting with 15% MeCN: water and increasing the amount of MeCN linearly over ten minutes to 90%. That concentration was held constant for 2 minutes before returning to initial conditions. During the entire analysis the flow rate was 0.9 mL/min.
Flow Injection Conditions: A 1:1 mixture of the water to MeCN (both with 0.2% formic acid added) was used to acquire the FIA data. The flow rate was set to 0.3 ml/min.
1H NMR
All 1H NMR spectra were acquired using a Bruker Instruments AMX-500 NMR spectrometer in the solvent given.